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Suppression of E-cadherin mediates gallotannin induced apoptosis in Hep G2 hepatocelluar carcinoma cells.

Han HJ, Kwon HY, Sohn EJ, Ko H, Kim B, Jung K, Lew JH, Kim SH - Int. J. Biol. Sci. (2014)

Bottom Line: Consistently, Immunofluorescence assay showed that E-cadherin or β-catenin expression was suppressed in a time dependent manner by gallotannin.Furthermore, silencing of E-cadherin by siRNA transfection method enhanced PAPR cleavage, caspase 3 activation and sub G1 population and attenuated the cell adhesion induced by gallotannin in Hep G2 cells.Overall, our findings demonstrate that the disruption of cell adhesion junction by suppression of E-cadherin mediates gallotannin enhanced apoptosis in Hep G2 liver cancer cells.

View Article: PubMed Central - PubMed

Affiliation: 2. Graduate School of East-West Medical Science, Kyung Hee University, Yongin 449-701, Republic of Korea.

ABSTRACT
Though gallotannin was known to have anti-oxidant and antitumor activity, the underlying antitumor mechanism of gallotannin still remains unclear. Thus, in the present study, antitumor mechanism of gallotannin was elucidated in hepatocellular carcinoma cells. Gallotannin significantly exerted cytotoxicity against Hep G2 and Chang hepatocellular carcinoma cells with the accumulation of the sub-G1 population and increase of terminal deoxynucleotidyltransferasedUTP nick end labeling (TUNEL) positive cells as an apoptotic feature. Also, gallotannin attenuated the expression of pro-caspase9, pro-caspase3, Bcl2 and integrin β1 and cleaved poly(ADP)-ribose polymerase (PARP) in Hep G2 and Chang cancer cells. Furthermore, gallotannin suppressed cell repair motility by wound healing assay and also inhibited cell adhesion in Hep G2 cells. Of note, gallotannin attenuated the expression of epithelial cadherin (E-cadherin) to form cell-cell adhesion from the early stage, and also beta-catenin at late phase in Hep G2 cells. Consistently, Immunofluorescence assay showed that E-cadherin or β-catenin expression was suppressed in a time dependent manner by gallotannin. Furthermore, silencing of E-cadherin by siRNA transfection method enhanced PAPR cleavage, caspase 3 activation and sub G1 population and attenuated the cell adhesion induced by gallotannin in Hep G2 cells. Overall, our findings demonstrate that the disruption of cell adhesion junction by suppression of E-cadherin mediates gallotannin enhanced apoptosis in Hep G2 liver cancer cells.

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Gallotannin attenuated the expression of E-cadherin, and beta-catenin in Hep G2 cells. (A) Effect of gallotannin on E-cadherin in Hep G2 cells in a concentration (0, 2, 4 μM) and time dependent manner (24, 48 or 72 h) by Western blotting (B) Effect of gallotannin on E-cadherin and beta-catenin in Hep G2 cells in time courses (0, 6, 18, 24, 48 h) by Western blotting. Effect of gallotannin on E-cadherin (C) or β-catenin (D) localization in Hep G2 cells in Hep G2 cells by Immunofluorescence assay. Hep G2 cells in the absence or presence of gallotannin (4μM) were fixed and immunostained with α-E-cadherin antibody. Nuclei were stained with DAPI. Scale bar, 40 μm.
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Figure 4: Gallotannin attenuated the expression of E-cadherin, and beta-catenin in Hep G2 cells. (A) Effect of gallotannin on E-cadherin in Hep G2 cells in a concentration (0, 2, 4 μM) and time dependent manner (24, 48 or 72 h) by Western blotting (B) Effect of gallotannin on E-cadherin and beta-catenin in Hep G2 cells in time courses (0, 6, 18, 24, 48 h) by Western blotting. Effect of gallotannin on E-cadherin (C) or β-catenin (D) localization in Hep G2 cells in Hep G2 cells by Immunofluorescence assay. Hep G2 cells in the absence or presence of gallotannin (4μM) were fixed and immunostained with α-E-cadherin antibody. Nuclei were stained with DAPI. Scale bar, 40 μm.

Mentions: There are evidences for the role of E-cadherin in apoptosis in cancer cells 20-23. In this regards, we examined the role of E-cadherin in gallotannin induced apoptosis in Hep G2 cells. As shown in Figure 4 A, gallotannin attenuated the expression of E-cadherin in a time dependent manner from 24 h after treatment, while the expression of beta catenin was effectively reduced in Hep G2 cells only at 72 h after treatment (Figure 4 A, B). We also confirmed that red color expression for E-cadherin around cell membrane or β-catenin was effectively attenuated in gallotannin treated Hep G2 cells compared to untreated control by immunofluorescence assay (Figure 4C or D).


Suppression of E-cadherin mediates gallotannin induced apoptosis in Hep G2 hepatocelluar carcinoma cells.

Han HJ, Kwon HY, Sohn EJ, Ko H, Kim B, Jung K, Lew JH, Kim SH - Int. J. Biol. Sci. (2014)

Gallotannin attenuated the expression of E-cadherin, and beta-catenin in Hep G2 cells. (A) Effect of gallotannin on E-cadherin in Hep G2 cells in a concentration (0, 2, 4 μM) and time dependent manner (24, 48 or 72 h) by Western blotting (B) Effect of gallotannin on E-cadherin and beta-catenin in Hep G2 cells in time courses (0, 6, 18, 24, 48 h) by Western blotting. Effect of gallotannin on E-cadherin (C) or β-catenin (D) localization in Hep G2 cells in Hep G2 cells by Immunofluorescence assay. Hep G2 cells in the absence or presence of gallotannin (4μM) were fixed and immunostained with α-E-cadherin antibody. Nuclei were stained with DAPI. Scale bar, 40 μm.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4007362&req=5

Figure 4: Gallotannin attenuated the expression of E-cadherin, and beta-catenin in Hep G2 cells. (A) Effect of gallotannin on E-cadherin in Hep G2 cells in a concentration (0, 2, 4 μM) and time dependent manner (24, 48 or 72 h) by Western blotting (B) Effect of gallotannin on E-cadherin and beta-catenin in Hep G2 cells in time courses (0, 6, 18, 24, 48 h) by Western blotting. Effect of gallotannin on E-cadherin (C) or β-catenin (D) localization in Hep G2 cells in Hep G2 cells by Immunofluorescence assay. Hep G2 cells in the absence or presence of gallotannin (4μM) were fixed and immunostained with α-E-cadherin antibody. Nuclei were stained with DAPI. Scale bar, 40 μm.
Mentions: There are evidences for the role of E-cadherin in apoptosis in cancer cells 20-23. In this regards, we examined the role of E-cadherin in gallotannin induced apoptosis in Hep G2 cells. As shown in Figure 4 A, gallotannin attenuated the expression of E-cadherin in a time dependent manner from 24 h after treatment, while the expression of beta catenin was effectively reduced in Hep G2 cells only at 72 h after treatment (Figure 4 A, B). We also confirmed that red color expression for E-cadherin around cell membrane or β-catenin was effectively attenuated in gallotannin treated Hep G2 cells compared to untreated control by immunofluorescence assay (Figure 4C or D).

Bottom Line: Consistently, Immunofluorescence assay showed that E-cadherin or β-catenin expression was suppressed in a time dependent manner by gallotannin.Furthermore, silencing of E-cadherin by siRNA transfection method enhanced PAPR cleavage, caspase 3 activation and sub G1 population and attenuated the cell adhesion induced by gallotannin in Hep G2 cells.Overall, our findings demonstrate that the disruption of cell adhesion junction by suppression of E-cadherin mediates gallotannin enhanced apoptosis in Hep G2 liver cancer cells.

View Article: PubMed Central - PubMed

Affiliation: 2. Graduate School of East-West Medical Science, Kyung Hee University, Yongin 449-701, Republic of Korea.

ABSTRACT
Though gallotannin was known to have anti-oxidant and antitumor activity, the underlying antitumor mechanism of gallotannin still remains unclear. Thus, in the present study, antitumor mechanism of gallotannin was elucidated in hepatocellular carcinoma cells. Gallotannin significantly exerted cytotoxicity against Hep G2 and Chang hepatocellular carcinoma cells with the accumulation of the sub-G1 population and increase of terminal deoxynucleotidyltransferasedUTP nick end labeling (TUNEL) positive cells as an apoptotic feature. Also, gallotannin attenuated the expression of pro-caspase9, pro-caspase3, Bcl2 and integrin β1 and cleaved poly(ADP)-ribose polymerase (PARP) in Hep G2 and Chang cancer cells. Furthermore, gallotannin suppressed cell repair motility by wound healing assay and also inhibited cell adhesion in Hep G2 cells. Of note, gallotannin attenuated the expression of epithelial cadherin (E-cadherin) to form cell-cell adhesion from the early stage, and also beta-catenin at late phase in Hep G2 cells. Consistently, Immunofluorescence assay showed that E-cadherin or β-catenin expression was suppressed in a time dependent manner by gallotannin. Furthermore, silencing of E-cadherin by siRNA transfection method enhanced PAPR cleavage, caspase 3 activation and sub G1 population and attenuated the cell adhesion induced by gallotannin in Hep G2 cells. Overall, our findings demonstrate that the disruption of cell adhesion junction by suppression of E-cadherin mediates gallotannin enhanced apoptosis in Hep G2 liver cancer cells.

Show MeSH
Related in: MedlinePlus